Mycoplasma is a common contaminant of cell and tissue culture samples. Infection is persistent, difficult to detect and diagnose, and very difficult to cure. The presence of Mycoplasma in infected cultures can change many of the cells' reactions, including altering cell growth rate, inducing morphological changes or cell transformation, and mimicking virus infection. A Mycoplasma-contaminated cell line is typically significantly influenced in every respect, and, thus, does not comply with Pharmacopoeia and FDA regulatory requirements. Therefore, there is an absolute requirement for routine, periodic assays for testing of possible contamination of all cell cultures used in manufacturing of pharmaceuticals.
Existing methods for identification of Mycoplasma contamination rely on traditional bacterial culturing on agarose plates, taking up to 28 days for results. Such a protracted time between sampling and results preclude efficient quality control (QC) checks, delay production approvals and limit sampling points for QC. PCR-based methods have the greatest potential to become a routine procedure for Mycoplasma testing for cell culture, tissue culture, food, environmental, agricultural, biopharmaceutical, and pharmaceutical QC. There remains, however, a need for a Mycoplasma detection assay that is, for example, rapid, sensitive, and specific for a variety of Mycoplasma species.